Chemistry: molecular biology and microbiology – Vector – per se
Reexamination Certificate
2006-07-04
2006-07-04
Devi, S. (Department: 1645)
Chemistry: molecular biology and microbiology
Vector, per se
C435S071100, C435S069600, C435S069300, C435S069100, C435S091400, C435S471000, C435S476000
Reexamination Certificate
active
07070989
ABSTRACT:
The present invention provides a recombinant plasmid vector comprising a kanamycin resistance gene, a promoter, an endoxylanase signal sequence, a nucleotide sequence coding for an oligopeptide consisting of 13 amino acids including 6 consecutive histidine residues, and a human granulocyte colony stimulating factor(hG-CSF) gene; anE. colitransformed with the said vector; and, a process for producing complete hG-CSF protein with high purity from the protein pool secreted by the said microorganism. In accordance with the invention, the hG-CSF protein can be prepared with high purity through rather simple process facilitating secretion of large amount of hG-CSF fusion protein into the periplasm, which does not require complicated processes such as solubilization and subsequent refolding required for isolation of the hG-CSF protein produced in cytoplasm as insoluble inclusion bodies by conventional techniques, thus, the hG-CSF protein can be widely used as an active ingredient in the development of supplementary agents for anticancer therapy.
REFERENCES:
Bong Hyun Chung, et al “Overproduction of Human Granulocyte-Colony Stimulating Factor Fused to the PelB Signal Peptide inEscherichia coli.” J Ferment Bioeng., 85(4) 443-446 (1998).
Julian Pérez-Pérez, et al. “DnaK/DnaJ Supplementation Improves the Periplasmic Production of Human Granulocyte-Colony Stimulating Factor inEscherichia coli.” Biochem. Biophys. Res. Commun., 210(2):524-529 (1995).
Jeong Ki-Jun
Lee Sang-Yup
Devi S.
Knobbe Martens Olson & Bear LLP
Korea Advanced Institute of Science and Technology
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